Isolation and Identification of Plant Growth-Promoting Bacteria Highly Effective in Suppressing Root Rot in Fava Beans.

In this study, 18 plant growth-promoting bacterial (PGPB) strains were isolated from the rhizosphere of both Red silk-cotton tree (Bombax malabaricum) and Chinese banyan (Ficus retusa). Culture morphology was observed and genotypic characterization was accomplished by sequencing partial 16S rRNA gene. Plant growth promotion traits and antagonistic activities of the strains against phytopathogenic fungi were evaluated. Among all strains, Bacillus thuringiensis (MN419208) exert the highest indole acetic acid (38 µg/ml), produced exopolysaccharides (587.2 µg/ml), and fixed nitrogen which in turn increased both fresh and dry weights of bean plants by 41.5% and 18.8%, respectively. In another greenhouse experiment studying the antifungal activities of seven strains and their co-culture against Rhizoctonia solani (LN735538), B. sonorensis MN419205, B. wiedmannii MN419207, B. subtilis MN419218, and the mixture of (MN419207) and (MN419208) reduced total damping off from 81.7% in control to 30%, 35%, 35%, and 38.3%, respectively, and reduced disease severity index from 33.3% to 20.5%, 22.5%, 14.2%, and 19.3% as well. Our data indicate that these strains are effective in promoting plant growth and in inhibiting R. solani infection nonetheless field experiments are needed to examine their effectiveness as a viable alternative to chemical fertilizers and conventional pesticides.

Optimization of methylene blue degradation by Aspergillus terreus YESM 3 using response surface methodology.

Synthetic dyes released from many industries cause pollution problems in aquatic environments affecting public health. The present study aimed to explore the potentiality of Aspergillus terreus YESM 3 (accession number LM653117) for colour removal of three different dyes: methylene blue (MB), malachite green (MG) and safranin (S). Results showed that the tolerance index of the studied fungus against tested dyes decreased in the order: methylene blue, safranin and malachite green. Removal of methylene blue colour was improved by using Box-Behnken design. Optimum condition for methylene blue biodegradation in Czapek Dox broth was achieved at pH 6, of 31.41 mg/L dye concentration and an inoculum of 5.7778 × 104 (conidia/mL) with biodegradation of 89.41%. Thus, a novel and eco-friendly system for the biodegradation of dyes using Box-Behnken design has been efficiently developed. Accordingly, A. terreus YESM 3 can be professionally used for bioremediation of methylene blue dye in wastewater and removal of environmental pollution.

Association of genetic polymorphisms of chemokines and their receptors with clearance or persistence of hepatitis C virus infection.

BACKGROUND: Polymorphisms of certain genes may have an effect on either persistence of infection or spontaneous clearance of hepatitis C virus (HCV). We hypothesized that one or more variants of chemokines (CCL2 and CCL5) and chemokine receptors (CC chemokine receptor type 2 [CCR2]) genes are associated with the susceptibility to HCV infection. METHODS: We recruited 1460 patients with chronic HCV (CHC), 108 subjects with spontaneous virus clearance (SVC) and 1446 individuals as a healthy control group. All were genotyped for single nucleotide polymorphisms: rs13900 C/T of CCL2, rs3817655 T/A of CCL5 and rs743660 G/A and rs1799864 G/A of CCR2 using allelic discrimination real-time PCR technique. RESULTS: The carriage of the A allele of CCR2 rs743660 was significantly higher in CHC compared to SVC (odds ratio [OR] 4.03) and to controls (1.42) and in controls compared to SVC (2.85) (all P < 0.01). Similarly, the A allele of CCR2 rs1799864 was significantly higher in the CHC group when compared with both SVC (1.97) and controls (2.13) (both P < 0.01), but the OR between controls and SVC was not significant (1.08, P = 0.723). Carriage of C allele of CCL2 rs13900 and the T allele of CCL5 rs3817655 were significantly higher in SVC group when compared with both CHC (OR = 0.19 and OR = 0.24, respectively) and control groups (OR = 0.65 and OR = 0.45, respectively [all P < 0.01]). CONCLUSIONS: Susceptibility to HCV infection is associated with A alleles of both (rs743660 and rs1799864 G/A) of CCR2 while spontaneous clearance of HCV is associated with the C allele of rs13900 of CCL2 and T allele of rs3817655 of CCL5.

Biodegradation of imidacloprid in liquid media by an isolated wastewater fungus Aspergillus terreus YESM3.

In the present study, a new fungal strain capable of imidacloprid degradation was isolated from agricultural wastewater drain. The fungal strain of YESM3 was identified as Aspergillus terreus based on ITS1-5.8S rDNA-ITS2 gene sequence by PCR amplification of a 500 bp sequence. Screening of A. terreus YESM3 to the insecticide imidacloprid tolerance was achieved by growing fungus in Czapek Dox agar for 6 days at 28°C. High values (1.13 and 0.94 cm cm-1) of tolerance index (TI) were recorded at 25 and 50 mg L-1 of imidacloprid, respectively in the presence and absence of sucrose. However, at 400 mg L-1 the fungus did not grow. Effects of the imidacloprid concentration, pH, and inoculum size on the biodegradation percentage were tested using Box-Behnken statistical design and the biodegradation was monitored by HPLC analysis at different time intervals. Box-Behnken results indicated that optimal conditions for biodegradation were at pH 4 and two fungal discs (10 mm diameter) in the presence of 61.2 mg L-1 of imidacloprid. A. terreus YESM3 strain was capable of degrading 85% of imidacloprid 25 mg L-1 in Czapek Dox broth medium at pH 4 and 28°C for 6 days under static conditions. In addition, after 20 days of inoculation, biodegradation recorded 96.23% of 25 mg L-1 imidacloprid. Degradation kinetics showed that the imidacloprid followed the first order kinetics with half-life (t50) of 1.532 day. Intermediate product identified as 6-chloronicotinic acid (6CNA) as one of the major metabolites during degradation of imidacloprid by using HPLC. Thus, A. terreus YESM3 showed a potential to reduce pollution by pesticides and toxicity in the effected environment. However, further studies should be conducted to understand the biodegradation mechanism of this pesticide in liquid media.

Application of artificial neural networks for enhancing Aspergillus flavipes lipase synthesis for green biodiesel production.

Biodiesel is a sustainable, and renewable alternative to fossil fuels that can be produced from various biological sources with the aid of lipases. This study developed a simple and novel fungal system for lipase biosynthesis to be used for catalyzing the oily residuals into biodiesel, employing the artificial neural network (ANN), and semi-solid-state fermentation (SSSF). Nigella sativa was selected among agro-industrial oily residuals as a substrate for lipase biosynthesis by Aspergillus flavipes MH47297. The effect of cultural humidity (X1), the surfactant; Brij 35 (X2), and inoculum density (X3) on lipase biosynthesis were researched based on the matrix of Box-Behnken design (BBD). The ANN together with a new fungal candidate and SSSF were then applied for the first time to model the biosynthesis process of lipase. The optimum predicted cultural conditions varied according to the model. The optimum predicted conditions were estimated separately by BBD (X1 = 5.8 ml water/g, X2 = 46.6 µl/g, and X3 = 62156610 spore/g) and ANN (X1 = 5.4 ml water/g, X2 = 54.2 µl/g, and X3 = 100000000 spore/g) models. Based on the modeling process, the response of lipase was calculated to be 214.95 (BBD) and 217.72 U (ANN), which revealed high consistency with the experimental lipase yield (209.13 ± 3.27 U for BBD, and 218 ± 2.01 U for ANN). Despite both models showing high accuracy, ANN was more accurate and surpassed the BBD model. Gas chromatography analysis showed that lipase successfully converted corn oil to biodiesel (29.5 mg/l).

The baculovirus promoter OpIE2 sequence has inhibitory effect on the activity of the cytomegalovirus (CMV) promoter in HeLa and HEK-293 T cells.

Understanding how promoters work in non-host cells is complex. Nonetheless, understanding this process is crucial while performing gene expression modulation studies. This study began with the process of constructing a shuttle vector with CMV and OpIE2 promoters in a tandem arrangement to achieve gene expression in both mammalian and insect cells, respectively. In this system, inhibitory regions in the 5' end of the OpIE2 insect viral promoter were found to be blocking the activity of the CMV promoter in mammalian cells. Initially, the OpIE2 promoter was cloned downstream of the CMV promoter and upstream of the EGFP reporter gene. After introducing the constructed shuttle vector to insect and mammalian cells, a significant drop in the CMV promoter activity in mammalian cells was observed. To enhance the CMV promoter activity, several modifications were made to the shuttle vector including site-directed mutagenesis to remove all ATG codons from the downstream promoter (OpIE2), separating the two promoters to eliminate the effect of transcription interference between them, and finally, identifying some inhibitory regions in the OpIE2 promoter sequence. When these inhibitory regions were removed, high expression levels in insect and mammalian cells were maintained. In conclusion, a shuttle vector was constructed that works efficiently in both mammalian and insect cell lines in the absence of baculovirus infection or gene expression. Moreover, the shuttle vector can be used as a platform to further study the reason for this inhibition, which may give new insights about transcription and promoters' mode of action in both insect and mammalian hosts.

Applications of Fusarium solani YMM20 in bioremediation of heavy metals via enhancing extracellular green synthesis of nanoparticles.

A novel technique for removing metals from water, by converting them into nanoparticles using the culture filtrate of Fusarium solani YMM20, followed by their removal by centrifugation was developed in this study. It is a promising, simple, and eco-friendly process for the bioremediation of wastewater. This method is built on the reduction of metal ions by cell-free fungal filtrate protein(s). The chitosan and chitosan nanoparticles were used as adsorbents for the removal of Cu2+ , Pb2+ , Co2+ , Cd2+ , Ni2+ , and Fe2+ , separately. In addition, fungal filtrate and chitosan nanoparticles loaded with fungal filtrate were used for the biosynthesis of metal nanoparticles of Cu2+ , Pb2+ , Co2+ , Cd2+ , Ni2+ , and Fe2+ , separately for their removal. The fungal filtrate approach has the highest metal removal (%) for most metals. Among the six metal ions, the fungal filtrate, chitosan nanoparticles loaded with fungal filtrate, chitosan, and chitosan nanoparticles showed the highest removal (%) for Pb2+ and the lowest removal (%) for Co2+ . Techniques such as transmission electron microscopy, scanning electron microscopy, and Zeta potential measurements were used to characterize the obtained nanoparticles. The average size of the spherical nanoparticles observed by transmission electron microscopy was from 18.938 ± 2.269 nm to 60.175 ± 4.973 nm.

Production, characterization and bio-emulsifying application of exopolysaccharides from Rhodotorula mucilaginosa YMM19.

Microbial exopolysaccharides (EPS) are high molecular weight polymers having different sugar residues. EPS have potential applications in different fields, such as medicine, food and environment. Therefore, there is a growing interest in production, characterization and application of EPS from different microorganisms. The present study designed to investigate the production and characterization of EPS from Rhodotorula mucilaginosa YMM19 isolated from Morus nigra L. fruits as well as to examine their potential emulsifying properties. Effect of NaCl concentration, incubation period and pH on the production of EPS was studied. The maximum EPS production by yeast was achieved at 10% NaCl (9741.84 mg/l). The best incubation time for production of EPS was 5 days. Production of EPS decreased under neutral condition and increased at acidic and alkaline condition. The structural feature of EPS was examined by FT-IR and NMR spectral analysis and confirmed the presence of glucose, glucopyranose and galactose. The isolated EPS showed higher emulsification capacity with emulsification activity of 71% and emulsifying index of 60%. The EPS gave strong emulsification for farnesol and was more effective than sodium dodecyl sulphate, a reference emulsifier, in enhancing the herbicidal activity of farnesol against Melilotus indicus under greenhouse condition. The results suggest that the EPS produced by YMM19 strain has a potential to be used as emulsifying agent in pesticide formulations.

Significantly reducing implant loss rates in immediate implant-based breast reconstruction: A protocol and completed audit of quality assurance.

INTRODUCTION: Immediate implant-based breast reconstruction (IBR) rates have increased considerably with the advent of acellular dermal matrices. Implant loss is a significant complication and is costly to patients and the NHS. National Mastectomy and Breast Reconstruction Audit and Implant-Based Breast Reconstruction Audit data have demonstrated national implant loss rate of 9% at 3 months. National Oncoplastic Guidelines for Best Practice cite a < 5% target. We aimed to reduce implant loss by introducing a protocol with pre-, intra- and post-operative interventions. METHODS: Audit of IBR at a single oncoplastic breast unit was commenced and implant loss at 3 months was recorded (May 2012-July 2014). Patients were identified from a prospectively maintained database, and case notes were examined by identifying factors associated with implant loss. A team involving microbiology, theatre staff, infection control and surgeons was established. A novel, evidence-based intervention bundle, including more than 25 protocol changes, was introduced. Prospective re-audit of IBR (April 2015-December 2017) was completed following introduction of the new protocol and implant loss was recorded at 3 months. RESULTS: The first retrospective audit of 77 reconstructions (54 patients) demonstrated 11 implant losses at 3 months (14%). Re-audit, post-intervention, comprised 129 reconstructions (106 patients) with no implant loss at 3 months. Fisher's exact analysis revealed statistically significant reduction in implant loss rate (P < 0.00001) following protocol introduction. CONCLUSIONS: Implant loss rate following IBR can be reduced to an exceptionally low level, well below national targets, by adhering to this evidence-based intervention bundle. Our protocol could improve outcomes nationally.

Cutaneous epithelioid angiomatous nodule: different views or interpretations in the analysis of ten new cases.

Classification schemes proposed for vascular lesions are the subjects of significant controversy. Cutaneous epithelioid angiomatous nodule (CEAN) was described in 2004, but there is no agreement as to whether this is a distinct entity or a type of either epithelioid hemangioma or angiolymphoid hyperplasia with eosinophilia. We present a typical case of CEAN and discuss nine other cases from our institution. We then provide two opposing viewpoints concerning its classification.

Preoperative estimation of cosmetic outcomes after immediate breast reconstruction with extended latissimus dorsi flap: A simple prediction model.

BACKGROUND: The extended latissimus dorsi (ELD) flap can provide adequate volume for breast reconstruction without an implant. The aim of this study was to identify a simple method to estimate preoperatively if the ELD flap would provide enough volume for breast reconstruction and good cosmetic outcome. The proposed model was based on correlating the preoperative body mass index (BMI) and breast cup size. METHODS: This retrospective cohort study included 64 patients who underwent immediate breast reconstruction with ELD at the National Cancer Institute, Cairo University, between January 2014 and January 2016. Preoperative breast cup size and patients' BMI were recorded. The primary end point was cosmetic outcome score assessed by three independent breast surgeons and the patients. Correlation analysis was performed between the preoperative factors and final cosmetic outcome. RESULTS: Sixty-four patients underwent immediate reconstruction following skin-sparing mastectomy. BMI range was 23-38 kg/m2 with a median of 28. All patients with cup size A had aesthetically pleasing results. As the breast cup size increased, the favorable cosmetic outcomes were noted with lower BMI. All patients with BMI higher than 33 had unfavorable results. CONCLUSION: Favorable cosmetic outcomes are expected after ELD reconstruction in patients with cup size A regardless of the BMI. As the breast cup size increases, favorable cosmetic outcomes are expected in patients with lower BMI than in those with higher BMI. At a BMI of 34, no favorable cosmetic outcomes are expected. The prediction model will be validated in a prospective study.

Urinary bisphenol A concentrations in relation to asthma in a sample of Egyptian children.

BACKGROUND: Bronchial asthma is one of the top disabling diseases in pediatrics. Limited research has been studied the association of the widely used plastic monomer bisphenol A (BPA) with childhood asthma. OBJECTIVE: To compare the levels of urinary BPA in asthmatic and control children and to investigate the implication of BPA among other risk factors for the development of asthma. SUBJECTS AND METHODS: This case-control study included 97 children (45 asthmatic and 52 healthy controls) aged 3-8 years. Asthmatic children were diagnosed according to Global initiative for asthma (GINA) guidelines. Sociodemographic factors were assessed and urinary levels of BPA were determined in spot urine samples using high-performance liquid chromatography. The contribution of BPA among predictors for developing asthma was studied in asthmatic children. RESULTS: Median total urinary BPA levels were significantly higher in asthmatic children than in control group (1.56 ng/mL in asthmatic children compared to 0.790 ng/mL in control group, p = 0.001). Children who had total urinary BPA levels >1.3 ng/mL were more likely to be asthmatic (odds ratio: 2.84, 95% confidence interval 1.22-6.59, p = 0.015). Multiple logistic regression analysis for predictors of asthma showed the importance of higher levels of BPA (>1.3 ng/mL) as a more significant predictor than passive smoking ( p = 0.006 for BPA categories vs. p = 0.049 for passive smoking). CONCLUSION: Association of higher levels of urinary BPA with the diagnosis of asthma in children may indicate the potential risk of BPA exposure in the precipitation of bronchial asthma. Further clinical and biochemical research are needed to clarify the proper mechanism explaining this association.

A trial to produce an anti-schistosomal vaccine using heterophyid antigens.

Heterophyid metacercariae, crude and partially purified heterophyid antigens were given prior to S. mansoni infections in an attempt to produce a potent antischistosomal vaccine. Three main groups of albino mice were used. Each group after receiving the appropriate heterophyid antigen prior to S. mansoni infection was studied parasitologically as regards worm load and tissue egg count and histopathologically as regards granulomata number, size and cellular constituents as well as immunologically by the indirect immunofluorescent test. The results showed a reduction in worm burden recoveries and egg load resulting in lesser number of granulomata and diminution in size of granulomata as well as acceleration in their reaction together with inhibition of fluorescence. These results were more obvious in the group infected one month post heterophyid infection, as well as in mice immunized with the highest dose of crude heterophyid antigen (200 micrograms protein). However, a low dose of 50 micrograms of partially purified fractionated antigen gave the most evident results.

Design and synthesis of azolopyrimidoquinolines, pyrimidoquinazolines as anti-oxidant, anti-inflammatory and analgesic activities.

The 5,10-dihydro-2-thioxo-pyrimido[4,5-b]quinolines (2a-c) and its oxidized form 3 were prepared and used as key intermediates for the synthesis of thiazolo[3',2':1,2]pyrimido[4,5-b]-quinolines (5a-c), isoxazolo[5'',4'':4',5']thiazolo[3',2':1,2]pyrimido[4,5-b]quinolines (6a-c), 4-chloro-2-methylthio-pyrimido[4,5-b]quinoline, its amino derivatives (19-21) and 10,11,12,13-tetrahydro-5H-quino[2',3':4,5]pyrimido[6,1-b]quinazoline (22). The newly synthesized compounds were characterized by IR, NMR (1H, 13C) and mass spectral studies. Representative of the synthesized compounds was tested and evaluated for anti-oxidant, anti-inflammatory and analgesic activities. Compounds 2a-c showed the highest inhibitory anti-oxidant activity either using erythrocyte hemolysis or ABTS methods. Compounds 2a, 10b, 16, and 17a manifested the best protective effect against DNA damage induced by bleomycin. Compounds 2c, 5a, 20a, 2a, and 2b exhibited a potent anti-inflammatory activity using carrageenan-induced paw edema test in rats.

EMAP-II-dependent lymphocyte killing is associated with hypoxia in colorectal cancer.

Endothelial-monocyte-activating polypeptide-II (EMAP-II) is a novel multifunctional polypeptide with proinflammatory activity. We have previously shown that the recombinant and native forms of EMAP-II can induce apoptosis in mitogen-stimulated lymphocytes, and that the release of this protein into the extracellular milieu is enhanced by hypoxia. We hypothesised that hypoxia may lead to death of tumour-infiltrating lymphocytes (TILs) via an EMAP-II-dependent mechanism, thereby assisting tumours to evade the immune system. In this study, we used immunohistochemistry to detect EMAP-II, active caspase-3 and cleaved Poly (ADP-ribose) Polymerase (PARP) as indicators of apoptosis in TILs, and carbonic anhydrase IX (CA IX) as a surrogate marker of hypoxia. EMAP-II expression is associated with regions of hypoxia, and furthermore there is a significant association between TILs apoptosis and the presence of hypoxia. Using a coculture model of colorectal cancer cell/lymphocyte interactions, we were also able to demonstrate lymphocyte apoptosis induced by tumour cells, with concomitant caspase-3 activity. Lymphocyte killing was enhanced by direct cell-cell contact, particularly by tumour cells exposed to hypoxic conditions. Our data support the hypothesis that hypoxia plays a role in immune evasion by tumour cells, through EMAP-II-dependent lymphocyte killing.

Formulation, acceptability, chemical composition and in vitro digestibility of novel snack food and meat ball analogue.

Potato purée and wheat flour were blended at a ratio of 7:1 w/w with 2 parts per weight of soybean protein concentrate (SPC) and/or faba bean-protein micellar mass (PMM). Two snack foods were formulated and deep fried. Spices, onion, parsley and chicken stock were added to the aforementioned two recipes to formulate meat ball analogues. Data revealed good acceptability of all products. Moreover, they can be considered as a good source for some essential amino acids; protein (approximately 26%); lipids (approximately 20%); carbohydrates (approximately 48%) and minerals. The in vitro digestibility ranged between 61.0% and 66.9%.

Selected nutritional, physical and sensory characteristics of pan and flat breads prepared from composite flours containing fababean.

Composite flour blends containing wheat (W), fababean (F), cottonseed and sesame flours were formulated to provide the FAO/WHO/UNU protein requirements for the 2-5 year old child, and evaluated in pan and flat bread applications. Water absorption of composite flour doughs was up to 35% greater than the control but gluten strength and slurry viscosities were markedly reduced. Loaf volume and specific volume of pan breads prepared from composite flours were 25-60% less than that of the control bread but flat breads tolerated the protein supplements extremely well. The W/F flat bread, containing 27% of fababean flour, received acceptable taste, texture and colour scores and was only slightly inferior to the control in puffing and layer separation. Additions of cottonseed or sesame flours to the W/F blend failed to improve sensory properties of the flat breads.

Formulation and characteristics of new recipes of Egyptian patti beans (Falafel).

New recipes of Falafel were formulated and evaluated regarding acceptability, proximate chemical composition and nutritive value. Soybean meal and/or cotyledons were used to substitute for faba bean at different levels (i.e. zero %, 20%, 40%, 60%, 80% and 100%. Data indicated that blends of faba bean (20%) and soybean meal and/or cotyledons (80%) were the most acceptable recipes compared to the control (100% faba bean) as well as the other substitution levels. Blending soybean with faba bean led to appreciable improvement in some essential amino acids. The in vitro digestibility for soybean blends were comparable to the control.